IL-1 beta and IL-6 in mouse parotid acinar cells: characterization of synthesis, storage, and release.
Synthesis, storage, and secretion of the proinflammatory cytokine interleukin-1 beta (IL-1 beta) and the anti-inflammatory cytokine IL-6 have not been established in normal exocrine gland secretory cells. Parotid glands and isolated acinar cells prepared from BALB/c mice were homogenized for RNA isolation and reverse transcription-polymerase chain reaction (RT-PCR). IL-1 beta and IL-6 enzyme-linked immunosorbent assays (ELISAs) were done on supernatants prepared from mouse parotid acinar cell (MPAC) preparations unstimulated or stimulated between 0 and 10 min with 10(-5) M norepinephrine at 37 degrees C. MPACs were fixed in paraformaldehyde, frozen sectioned for light and electron microscopy, and labeled with antibodies to IL-1 beta and IL-6. Mouse specific riboprobes to IL-1 and IL-6 were used for in situ hybridization. RT-PCR yielded the expected IL-1 (336-bp) and IL-6 (614-bp) mRNA products. By ELISA, stimulated MPACs showed a significant increase in IL-1 beta (P < 0.03) and IL-6 (P < 0.01) release into supernatants by 10 min that paralleled the time course of amylase release. In situ hybridization showed the presence of transcripts for IL-1 and IL-6 in glandular epithelial cells. Gold-labeled IL-1 beta and IL-6 were significantly higher (P < 0.01) in granules than in the nucleus and cytoplasm. This study shows that MPACs synthesize IL-1 beta and IL-6 and release these cytokines from their granules after alpha- and beta-adrenergic stimulation.
Tanda
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N
Department of Pathology, Beth Israel Deaconess Medical Center, Boston, Massachusetts, USA.
Ohyama
H
H
Yamakawa
M
M
Ericsson
M
M
Tsuji
T
T
McBride
J
J
Elovic
A
A
Wong
D T
DT
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D000681:Amylases / Q000378:metabolism; D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D017403:In Situ Hybridization; D007375:Interleukin-1 / Q000096:biosynthesis / Q000378:metabolism*; D015850:Interleukin-6 / Q000096:biosynthesis / Q000378:metabolism*; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D008297:Male; D051379:Mice; D008854:Microscopy, Electron; D016253:Microscopy, Immunoelectron; D009638:Norepinephrine / Q000494:pharmacology; D015388:Organelles / Q000187:drug effects / Q000276:immunology / Q000648:ultrastructure; D010306:Parotid Gland / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D016133:Polymerase Chain Reaction; D014158:Transcription, Genetic / Q000187:drug effects
D017931:DNA Primers; D007375:Interleukin-1; D015850:Interleukin-6; D007770:L-Lactate Dehydrogenase; D000681:Amylases; D009638:Norepinephrine